Methods of using crystals of beta amyloid cleaving enzyme (BACE)

Abstract

Methods of using monoclinic crystals of human beta-secretase (BACE) having unit cell dimensions of a, b, and c, wherein a is about 81±20 Å to about 101 Å, b is 103±20 Å, c is 100±20 Å, and α=γ=90°, and β is 105°±10° for crystals of symmetry P2 1 and a=73.1, b=105.1, c=50.5 Å and β is 94.8° for crystals of C2 symmetry in drug screening assays comprising selecting a potential modifier by adding the potential modifiers to an aqueous mixture of the crystal and detecting a measure of binding, such that the potential modifier that binds is selected as a potential drug.

1. A method of using an isolated crystal of beta secretase (BACE) of SEQ ID NO:3 having space group symmetry C2, unit cell dimensions of a, b, and c, wherein a is about 53 Å to about 93 Å, b is about 85 Å to about 125 Å, and c is about 40 Å to about 60 Å; α=γ=90°, β is about 85° to about 105°;
 selecting a potential modifier by performing rational drug design with a three-dimensional structure determined for the crystal, wherein selecting is performed in conjunction with computer modeling; 
 adding the potential modifier to a binding assay; and 
 detecting a measure of binding, wherein the potential modifier that binds is selected as a potential drug. 
 
     
     
       2. A method of using an isolated co-crystal of beta secretase (BACE) of SEQ ID NO:3 having space group symmetry C2, unit cell dimensions of a, b, and c, wherein a is about 53 Å to about 93 Å, b is about 85 Å to about 125 Å, and c is about 40 Å to about 60 Å; α=γ=90°, β is about 85° to about 105°;
 selecting a potential modifier by performing rational drug design with a three-dimensional structure determined for the crystal, wherein selecting is performed in conjunction with computer modeling; 
 adding the potential modifier to a binding assay; and 
 detecting a measure of binding, wherein the potential modifier that binds is selected as a potential drug. 
 
     
     
       3. The method of  claim 2  wherein the exposing comprises soaking. 
     
     
       4. The method of  claim 2  wherein the samples include a variety of different functional groups. 
     
     
       5. The method of  claim 3 , wherein the crystal of BACE wherein a is 73.1 Å, b=105.1 Å, c=50.5 Å, α=γ=90°, and β=94.8°. 
     
     
       6. The method of  claim 2  further comprising identifying the ligand that forms the ligand-BACE molecular complex. 
     
     
       7. The method of  claim 6  wherein one of the determining and identifying comprises collecting x-ray diffraction data. 
     
     
       8. The method of  claim 7  wherein one of the determining and identifying comprises calculating an electron density function. 
     
     
       9. A method of using an isolated crystal of beta secretase (BACE) of SEQ ID NO:1 or SEQ ID NO:2 having space group symmetry P21, unit cell dimensions of a, b, and c, wherein a is about 61 Å to about 101 Å, b is about 83 Å to about 123 Å, and c is about 80 Å to about 120 Å; α=γ=90°, β is about 95° to about 115°;
 selecting a potential modifier by performing rational drug design with a three-dimensional structure determined for the crystal, wherein selecting is performed in conjunction with computer modeling; 
 adding the potential modifier to a binding assay; and 
 detecting a measure of binding, wherein the potential modifier that binds is selected as a potential drug. 
 
     
     
       10. A method of using an isolated co-crystal of beta secretase (BACE) of SEQ ID NO:1 or SEQ ID NO :2 having space group symmetry P2 1 , unit cell dimensions of a, b, and c, wherein a is about 61 Å to about 101 Å, b is about 83 Å to about 123 Å, and c is about 80 Å to about 120 A; α=γ=90°, β is about 95° to about 115°;
 selecting a potential modifier by performing rational drug design with a three-dimensional structure determined for the crystal, wherein selecting is performed in conjunction with computer modeling; adding the potential modifier to a binding assay; and 
 detecting a measure of binding, wherein the potential modifier that binds is selected as a potential drug. 
 
     
     
       11. The method of  claim 10  wherein the exposing comprises soaking. 
     
     
       12. The method of  claim 11  wherein the samples include a variety of different functional groups. 
     
     
       13. The method of  claim 10  further comprising identifying the ligand that forms the ligand-BACE molecular complex. 
     
     
       14. The method of  claim 13  wherein one of the determining and identifying comprises collecting x-ray diffraction data. 
     
     
       15. The method of  claim 13  wherein one of the determining and identifying comprises calculating an electron density function.